The ACAT1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered to disrupt the ACAT1 gene locus through targeted genome editing. This polyclonal product provides a heterogeneous population of cells with ACAT1 loss-of-function, enabling robust functional genomics studies without the clonal selection biases associated with single-cell derived lines. It serves as a versatile tool for investigating ACAT1-dependent metabolic processes in a cancer-relevant epithelial background.
The HT29 host cell line originates from a female patient with colorectal adenocarcinoma and exhibits adherent, epithelial-like growth characteristics. HT29 cells are widely used as an in vitro model for intestinal epithelial cell differentiation and colorectal cancer progression. Their capacity to undergo enterocytic differentiation under appropriate culture conditions makes them particularly suited for studying metabolic adaptations in the intestinal epithelium and their dysregulation in tumorigenesis.
ACAT1 encodes mitochondrial acetoacetyl-CoA thiolase, a homotetrameric enzyme that catalyzes the reversible condensation of two acetyl-CoA molecules to acetoacetyl-CoA, a critical step in ketone body production and isoleucine degradation. Regulated by PPARA, HNF4A, and insulin/glucagon signaling, ACAT1 interacts with HMGCS2 in the ketogenic pathway and influences the acetyl-CoA pool and synthesis of ketone bodies acetoacetate and ??-hydroxybutyrate. Its disruption impairs ketogenesis and branched-chain amino acid catabolism, with downstream effects on energy homeostasis.
In the context of HT29 colorectal cancer cells, ACAT1 knockout provides a physiologically relevant model to dissect the role of mitochondrial ketogenesis in cancer cell metabolism. Colorectal tumors often exhibit altered metabolic profiles, and ACAT1-mediated ketone body production may contribute to metabolic flexibility and stress responses. The HT29 background offers a platform to examine how ACAT1 deficiency impacts proliferation, survival, and metabolic reprogramming under nutrient-limited conditions, such as those encountered in the tumor microenvironment. This model can help elucidate the interplay between ketone body metabolism and oncogenic signaling in intestinal epithelial cells.
These polyclonal knockout cells are suitable for metabolic profiling of colorectal cancer, investigation of ketogenesis in intestinal epithelium, and drug screening for ketolytic defects. Researchers can employ techniques such as Western blotting, RT-qPCR, enzyme activity assays, ketone body quantification, Seahorse metabolic flux analysis, and proliferation assays to characterize ACAT1 loss-of-function. This model enables dissection of metabolic dependencies in colorectal cancer and identification of therapeutic targets. For further details, please contact Ascent Research.
